黄芪甲苷后处理对乳鼠心肌细胞缺氧复氧损伤的作用研究

目的:观察黄芪甲苷(AstragalosideⅣ,AsⅣ)后处理对缺氧复氧损伤(simulated ischemia reperfusion injury,SI/RI)的SD乳鼠心肌细胞是否具有保护作用。方法:将乳鼠原代心肌细胞平均分为五组,即空白对照组(Control)、缺氧复氧处理组(SI/RI)、黄芪甲苷预处理(5,10、20μM)+SI/RI组(AsIV+SI/RI)。各组细胞经处理后,四氮唑溴盐比色法(MTT)检测各组细胞存活率;TUNEL染色法测定各组细胞凋亡率;SOD测试盒检测培养液中超氧化物歧化酶(SOD)含量,总嘌呤氧化酶(XOD)测试盒检测丙二醛(MDA)含量。Western blot法检测各组细胞抗凋亡蛋白Bcl-2和促凋亡蛋白Caspase-3的表达。结果:与空白组相比,缺氧复氧损伤组细胞活力显著下降(P0.05),凋亡率显著上升(P0.05),其培养液中SOD水平显著降低(P0.05),MDA水平显著升高。而不同浓度AsⅣ后处理组的心肌细胞存活率显著上升,凋亡率显著下降,培养液中SOD水平显著上升,MDA水平显著下降(P0.05),且呈浓度呈依赖性。Western blot结果显示AsⅣ后处理组细胞中的Bcl-2表达明显上升,Caspase-3明显下降。结论:黄芪甲苷后处理对缺氧复氧诱导的乳鼠心肌细胞损伤具有显著的保护作用,能够显著上调抗凋亡蛋白Bcl-2的表达,下调促凋亡蛋白Caspase-3的表达。

Effects of Astragaloside IV on Neonatal Rat Cardiomyocytes Induced by Simulated Ischemia Reperfusion Injury*

Objective： To investigate the protective effects of post-treatment with Astragaloside Ⅳ（AsⅣ） on neonatal rat cardiom- yocytes induced by simulated ischemia reperfusion injury （SI/RI）. Methods： The cardiomyocytes were divided into 5 groups： Control, SL/RI, AsⅣ （5,10,20μM）＋SL/RI. After each group been treated, MTT method was used to determine the survival rate of cells; TUNEL used to detect the apoptotic rate of cells; The special kits were used to detect the levels of superpxide dismutase （SOD）and malondialdehyde （MDA） in the culture medium. Western blot was used to detect the expression of Bcl-2 and Caspase-3 protein. Results： Comparing with the control group, the cell survival rate and the apoptotic rate increased significantly, while the SOD level in the culture medium decreased significantly and the MDA level increased significantly. The above parameters of ceils were improved significantly when treated by As Ⅳ on concentration - dependent manner. Westren blot showed the expression of Bcl-2 protein increased significantly and the expression of Casepase-3 decreased significantly after being treated with As Ⅳ. Conclusion： Astragaloside Ⅳ has the protective effect on the neonatal rat cardiomyocytes induced by SI/RI, up-regulated the expression of Bcl-2 and down-regulated Caspase-3. Hence, Astragaloside Ⅳ may be a potential new drug in treatment of myocardial ischemia reperfusion injury.